• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.323-330
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• 1998

Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0～9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.

• Proceedings of the KAIS Fall Conference
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• 2010.11a
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• pp.495-499
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• 2010

The biocatalytic capture of $CO_2$, and its precipitationas $CaCO_3$, over bovine carbonic anhydrase (BCA) immobilized on a pore-expanded SBA-15 support was investigated. SBA-15 was synthesized using TMB as a pore expander, and the resulting porous silica was characterized by XRD, BET, IR, and FE-SEM analysis. BCA was immobilized on SBA-15 through various approaches, including covalent attachment (BCA-CA), adsorption (BCA-ADS), and cross-linked enzyme aggregation (BCA-CLEA). The immobilization of BCA on SBA-15 was confirmed by the presence of zinc metal in the EDXS analysis. The effects of pH, temperature, storage stability, and reusability on the biocatalytic performance of BCA were characterized by examining para-nitrophenyl acetate (p-NPA) hydrolysis. The $K_{cat}/K_m$ values for p-NPA hydrolysis were 740.05, 660.62, and $680.11M^{-1}s^{-1}$, respectively, where as $K_{cat}/K_m$ for free BCA was $873.76M^{-1}s^{-1}$. The amount of $CaCO_3$ precipitate was measured quantitatively using anion-selective electrode and was found to be 12.41, 11.82, or 11.28 mg $CaCO_3$/mg for BCA-CLEA, BCA-ADS, or BCA-CA, respectively. The present results indicate that the immobilized BCA-CLEA, BCA-ADS, and BCA-CA are green materials, and are tunable, reusable, and promising biocatalysts for $CO_2$ sequestration.

• Journal of the Korean Applied Science and Technology
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• v.29 no.1
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• pp.19-32
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• 2012

Pharmaceutical use accounts for a great part of articles and papers on crosslinking of polymers. Crosslinking of polymers used for tissue engineering and drug delivery respects non-cytotoxicity and in situ gelling. The crosslinking of polymers is aimed not only at the improvement of modulus, chemical resistance, and thermal resistance, but also at endowing them with such functions as metal adsorption, antifouling, and ion exchange via crosslinked segments. Smart polymers responding to environmental change, and cosslinking mediated by light, enzyme, natural compound and in aqueous medium in consideration of environment are being studied. Developing new polymeric materials is essential along with the pharmaceutics aiming at the longevity of 120 years old. Functionalization and property adjustment of polymers through crosslinking will be done more delicately. Hydrogels will be focused on injectable and in situ gel forming. In the coating industry crosslinking system with low non-toxicity and low energy consumption will be developed in consideration of workers and environment.

• The Journal of Korean Society of Virology
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• v.30 no.1
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• pp.61-70
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• 2000

Standard plaque assay using agarose-overlay has long been used for titration of many infectious virus particle. Plaque assay for the titration of varicella-zoster virus and its live vaccine requires three intermittent agarose overlay to visualize plaques. Overall procedure of the assay takes at least nine days from virus inoculation and microbe contamination including fungi is frequently accompanied during incubation period. We studied whether an immunofocus assay in conjunction with peroxidase-mediated immunohistochemical reaction may replace the standard plaque assay for the virus titration by comparing the two methods. A linear relationship was observed between number of foci and virus dilution. The number of foci in a given dilution of virus appeared a little higher than counted plaques formed in standard plaque assay. Independent titration results obtained from two assay methods for a given dilution of virus demonstrated a strong correlation ($r^2=0.99$). Foci of virus infected cells as revealed by the enzyme reaction could be counted either 4 days post-infection (p.i.) under low magnification (40X) microscopy, or 6 days p.i. by naked eye observation. Larger size of cell cuture plate, virus adsorption at $35^{\circ}C$, and 10% FBS in diluent appeared to be better conditions for the assay. Immunofocus assay will be an effective and dependable titration method for varicella-zoster virus and its live vaccine in place of the standard plaque assay in respect to accuracy, costs, and experimental convenience.

• Journal of Microbiology
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• v.45 no.5
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• pp.409-417
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• 2007

Burkholderia sp. HY-10 isolated from the digestive tracts of the longicorn beetle, Prionus insularis, produced an extracellular lipase with a molecular weight of 33.5 kDa estimated by SDS-PAGE. The lipase was purified from the culture supernatant to near electrophoretic homogenity by a one-step adsorption-desorption procedure using a polypropylene matrix followed by a concentration step. The purified lipase exhibited highest activities at pH 8.5 and $60^{\circ}C$. A broad range of lipase substrates, from $C_4\;to\;C_{18}$ p-nitrophenyl esters, were hydrolyzed efficiently by the lipase. The most efficient substrate was p-nitrophenyl caproate ($C_6$). A 2485 bp DNA fragment was isolated by PCR amplification and chromosomal walking which encoded two polypeptides of 364 and 346 amino acids, identified as a lipase and a lipase foldase, respectively. The N-terminal amino acid sequence of the purified lipase and nucleotide sequence analysis predicted that the precursor lipase was proteolytically modified through the secretion step and produced a catalytically active 33.5 kDa protein. The deduced amino acid sequence for the lipase shared extensive similarity with those of the lipase family 1.2 of lipases from other bacteria. The deduced amino acid sequence contained two Cystein residues forming a disulfide bond in the molecule and three, well-conserved amino acid residues, $Ser^{131},\;His^{330},\;and\;Asp^{308}$, which composed the catalytic triad of the enzyme.

• Fashion & Textile Research Journal
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• v.22 no.4
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• pp.534-539
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• 2020

Experimental research is needed to provide information on the removal of bloodstains since washing clothes contaminated with blood is necessary for medical related fields (such as ambulance workers and doctors) as well as for women of childbearing age. This study investigated efficient washing conditions for the removal of bloodstains with a focus on washing temperature, fiber type and blood ageing time. Polyester/cotton fabric showed the highest detergency from among three fabrics that were influenced by the composition of the fiber and the structure of the yarn and fabric. When examining the effect of detergent, it was concluded that the alkalinity over pH 10 was essential to remove bloodstains and that auxiliary agents such as soil antiredeposition agents and bleach had a significant effect on the removal of bloodstains. Washing temperature showed the highest detergency at 20℃ due to the activity of the enzyme without the denaturalization of blood. Blood-ageing influenced detergency by inducing changes in the adsorption area and chemical bond. A combination of methods such as quick removal after contamination, use of alkaline detergents including soil antiredeposition agents and bleach, and low-temperature washing could help remove bloodstains.

• Applied Biological Chemistry
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• v.35 no.5
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• pp.410-414
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• 1992

Endoinulase from Streptomyces sp. S56 was immobilized by adsorption on DEAE-cellulose in 0.01 M citrate-sodium phosphate buffer, pH 6.0 and the properties of immobilized and free enzymes were investigated. The immobilized enzyme preparation, having 40 inulase activity units per dried matrix, revealed the maximal activity at $pH\;4.5{\sim}5.5$ and $55{\sim}60^{\circ}C$ and were most stable at pH 6 and 45^{\circ}C$. The immobilization caused a drop in optimum pH and affinity toward inulin, a slight increase in optimum temperature, an important increase in thermal stability and maximum reaction velocity. The immobilized endoinulase hydrolyzed the tuber extract of jerusalem artichoke and inulin, mainly into fructose and inulobise, degrading 63 and 78% of the total sugar respectively, within 48 hrs in batch reactor.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1088-1097
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• 2021

Grouper nervous necrosis virus (GNNV) infection causes mass grouper mortality, leading to substantial economic loss in Taiwan. Traditional methods of controlling GNNV infections involve the challenge of controlling disinfectant doses; low doses are ineffective, whereas high doses may cause environmental damage. Identifying potential methods to safely control GNNV infection to prevent viral outbreaks is essential. We engineered a virus-binding bacterium expressing a myxovirus resistance (Mx) protein on its surface for GNNV removal from phosphate-buffered saline (PBS), thus increasing the survival of grouper fin (GF-1) cells. We fused the grouper Mx protein (which recognizes and binds to the coat protein of GNNV) to the C-terminus of outer membrane lipoprotein A (lpp-Mx) and to the N-terminus of a bacterial autotransporter adhesin (Mx-AIDA); these constructs were expressed on the surfaces of Escherichia coli BL21 (BL21/lpp-Mx and BL21/Mx-AIDA). We examined bacterial surface expression capacity and GNNV binding activity through enzyme-linked immunosorbent assay; we also evaluated the GNNV removal efficacy of the bacteria and viral cytotoxicity after bacterial adsorption treatment. Although both constructs were successfully expressed, only BL21/lpp-Mx exhibited GNNV binding activity; BL21/lpp-Mx cells removed GNNV and protected GF-1 cells from GNNV infection more efficiently. Moreover, salinity affected the GNNV removal efficacy of BL21/lpp-Mx. Thus, our GNNV-binding bacterium is an efficient microparticle for removing GNNV from 10‰ brackish water and for preventing GNNV infection in groupers.

• The Korean Journal of Mycology
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• v.31 no.2
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• pp.59-66
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• 2003

Laccase produced by Pycnoporus cinnabarinus SCH-3 isolated from Korea was partially purified using ultrafiltration, anion exchange chromatography and affinity chromatography, The laccase was produced as the predominant extracellular phenoloxidase during primary metabolism. Neither lignin peroxidase nor manganese-dependent peroxidase were detected in the culture fluid. In order to examine the effect of inducers in laccase production, 2,5-xylidine was added in the culture of Pycnoporus cinnabarinus SCH-3. Addition of 2,5-xylidine enhanced 25-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 9%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to be $44.4{\mu}M\;and\;56.0{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $60{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 10 min at $80{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the P. cinnabarinus SCH-3 laccase showed 94% of homology to the N-terminal sequences of laccases from P. cinnabarinus PB and P. coccineus.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.1
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• pp.97-104
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• 2002

Protein hydrolysate was prepared as a natural flavor stock from the sharp toothed eel (Muraenesox cinereus) mince using com-mercially available proteolytic enzymes, Alcalase, Neutrase, Protamex, and Flavourzyme. A 6 hr hydrolysis of mince, to which water of the equal weight of the mince was added, with $2\%$ (w/w, protein weight) Elavourzyme at $50^{\circ}$ yielded a hydrolysate of the highest acceptability. Removing the access lipid in liquified hydrolysate (not dehydrated) after enzyme hydrolysis, five times repetitive extraction using n-hexane (liquified hydrolysate : n-hexane=4 : 1, v/v) was effective, resulting in less than $1\%$ lipid content of the dehydrated-hydrolysate. The amino acid composition of the hydrolysate (prepared with Flavourzyme) was similar to that of the starting material. Hydrolysis led to an increase in concentration of not only total free amino acid but also free amino acid such as serine, glutamic acid, alanine, and methionine responsible for umami taste, especially up to about 40 times for methionine. Major free amino acids in amount were leucine, phenylalanine, valine, alanine, and isoleucine and they comprised about half of the total free amino acids, Moisture adsorption, fat adsorption, emulsifying capacity, and foaming capacity of the hydrolysate were 870.1 $\pm$ $7.9\%$, 352.0$\pm$ $5.3\%$, 50.3 $\pm$ $1.2\%$, and $87.5\pm$ $2.5\%$, respectively, and solubility was 83$\~$$84\%$ at acid pH range of 2$\~$4.